Recombinant Mouse Complement Component C3d Protein, CF

Catalog #: 2655-C3 Datasheet / COA / SDS

His-tag

Catalog #	Availability	Size / Price	Qty
2655-C3-050

Recombinant Mouse Complement Component C3d Protein Binding Activity

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All Complement Component C3d Proteins and Enzymes

Product Details

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Complement Component C3d Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Mouse Complement Component C3d His tag (Catalog # 2655-C3) is immobilized at 2 µg/mL (100 µL/well), (Catalog # 5980-CH) binds with an ED₅₀ of 1-6 ng/mL.

Source

Mouse myeloma cell line, NS0-derived mouse Complement Component C3d protein

IGS	Mouse C3d His1002-Arg1303 (Cys1010Ser) Accession # P01027	GSGGG GSGGG GSGS	Mouse C3d His1002-Arg1303 (Cys1010Ser) Accession # P01027	GSGGG GSGGG GSGS	Mouse C3d His1002-Arg1303 (Cys1010Ser) Accession # P01027	LE	HHHHHHHHHH
N-terminus							C-terminus

Accession #

P01027

N-terminal Sequence
Analysis

Ile-Gly-Ser-His1002

Predicted Molecular Mass

105 kDa

SDS-PAGE

80-95 kDa, reducing conditions

Product Datasheets

2655-C3

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2655-C3

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Recombinant Mouse Complement Component C3d His tag (Catalog # 2655-C3) is immobilized at 2 µg/mL, 100 µL/well, (Catalog # 5980-CH) binds with an ED₅₀ of 1-6 ng/mL.

SDS-PAGE

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2 μg/lane of Recombinant Mouse Complement Component C3d His tag (Catalog # 2655-C3) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands 80-95 kDa.

Reconstitution Calculator

Background: Complement Component C3d

Component C3 plays key role in activation of complement system. Proteolytic cleavage of full-length mouse C3 generates C3 beta (aa 25-666) and C3 alpha (aa 671-1663). C3 alpha is activated by proteolytic enzyme ‘C3 convertase', generating the small C3a peptide (aa 671-748), also called anaphylatoxin, and the large C3b alpha' chain (aa 749-1663). After activation, C3a functions as a mediator of local inflammatory process, while an internal thioester on C3b is exposed to mediate covalent attachment of C3b to foreign pathogens (1). C3b is further cleaved by several proteases to smaller fragments including C3g (aa 955-1001), C3d (aa 1002-1303), C3f (aa 1304-1320) and two C3c fragments (aa 749-954 and aa 1321-1663). The C3d fragment is a complement receptor 2 (CR2)-binding protein.The native C3d has an alpha -alpha barrel structure (2) which contains an isoglutamyl cysteine thioester cross link between Cys1010 and Gln1013 and a disulfide bond between Cys1101 and Cys1158, which allows C3d to covalently attach to cell surface molecules (2). In this recombinant mouse C3d protein, the Cys1010Ser mutation is introduced to ensure correct disulfide bonding between Cys1101 and Cys1158, since Cys1010 contributes the thiol moiety of the thioester, which would only be expected to form in native C3 (1). The biological function of this mutant C3d and its binding to CR2 is comparable to serum derived C3 fragment (2). Mouse C3d fragment shares 94% aa identity with rat c3d and 84% aa identity with human c3d. The interaction of C3d with CR2 enhances antibody responses by reducing the threshold for B cell activation (3). It also facilitates retention and internalization of antigens (4). C3d regulates immune checkpoint blockade and enhances antitumor immunity (5).

References